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Journal of Virology, January 2000, p. 110-116, Vol. 74, No. 1
0022-538X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

An 80-Kilodalton Protein That Binds to the Pre-S1 Domain of Hepatitis B Virus

Chun Jeih Ryu,1 Dae-Yeon Cho,1 Philippe Gripon,2 Hee Sun Kim,1 Christiane Guguen-Guillouzo,2 and Hyo Jeong Hong1,*

Antibody Engineering Research Unit, Korea Research Institute of Bioscience and Biotechnology, Yuseong, Taejon 305-600, Korea,1 and Hepatologique U49, Institut National de la Santé et de la Recherche Médicale, Hôpital de Pontchaillou, 35033 Rennes Cedex, France2

Received 8 June 1999/Accepted 22 September 1999

It has been suggested that hepatitis B virus (HBV) binds to a receptor on the plasma membrane of human hepatocytes via the pre-S1 domain of the large envelope protein as an initial step in HBV infection. However, the nature of the receptor remains controversial. In an attempt to identify a cell surface receptor for HBV, purified recombinant fusion protein of the pre-S1 domain of HBV with glutathione S-transferase (GST), expressed in Escherichia coli, was used as a ligand. The surface of human hepatocytes or HepG2 cells was biotinylated, and the cell lysate (precleared lysate) which did not bind to GST and glutathione-Sepharose beads was used as a source of receptor molecules. The precleared lysate of the biotinylated cells was incubated with the GST-pre-S1 fusion protein, and the bound proteins were visualized by Western blotting and enhanced chemiluminescence. An approximately 80-kDa protein (p80) was shown to bind specifically to the pre-S1 domain of the fusion protein. The receptor binding assay using serially or internally deleted segments of pre-S1 showed that amino acid residues 12 to 20 and 82 to 90 are essential for the binding of pre-S1 to p80. p80 also bound specifically to the pre-S1 of native HBV particles. Analysis of the tissue and species specificity of p80 expression in several available human primary cultures and cell lines of different tissue origin showed that p80 expression is not restricted to human hepatocytes. Taken together the results suggest that p80 may be a component of the viral entry machinery.

* Corresponding author. Mailing address: Antibody Engineering Research Unit, Korea Research Institute of Bioscience and Biotechnology, P.O. Box 115, Yuseong, Taejon 305-600, Korea. Phone: 82-42-860-4122. Fax: 82-42-860-4597. E-mail: hjhong{at}kribb4680.kribb.re.kr.

Journal of Virology, January 2000, p. 110-116, Vol. 74, No. 1
0022-538X/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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